Piatigorsky J, Horwitz J, Kuwabara T, Cutress C E
Laboratory of Molecular and Developmental Biology, National Eye Institute, Bethesda, Maryland 20892.
J Comp Physiol A. 1989 Feb;164(5):577-87. doi: 10.1007/BF00614500.
The ultrastructure and major soluble proteins of the transparent eye lens of two cubomedusan jellyfish, Tripedalia cystophora and Carybdea marsupialis, have been examined. Each species has two complex eyes (one large and one small) on four sensory structures called rhopalia. The lenses consist of closely spaced cells with few organelles. The lens is situated next to the retina, with only an acellular layer separating it from the photoreceptors. SDS-PAGE showed that the large lens of C. marsupialis has only two crystallin polypeptide bands (with molecular masses of approximately 20,000 and 35,000 daltons), while that of T. cystophora has three bands (two with a molecular mass near 20,000 daltons and one with a molecular mass near 35,000 daltons). Interestingly, the small lens of T. cystophora appears to be markedly deficient in or lack the lower molecular weight proteins. The crystallins behaved as monomeric proteins by FPLC and showed no immunological reaction with antisera of the major squid crystallin, chicken delta-crystallin or mouse gamma-crystallin in western immunoblots. Very weak reactions were found with antimouse alpha- and beta-crystallin sera. The 35,000 dalton crystallin of T. cystophora was purified and called J1-crystallin. It contained relatively high leucine (13%) and tyrosine (9%) and low methionine (2%). Several tryptic peptides were sequenced. Weak sequence similarities were found with alpha- and beta-crystallins, which may account for some of the apparent weak immunological cross-reactivity with these vertebrate crystallins. A polyclonal antiserum made in rabbits from a synthetic peptide of J1-crystallin reacted strongly with J1-crystallin of T. cystophora and C. marsupialis in immunoblots; by contrast, no reaction was obtained with the lower molecular weight crystallins from these jellyfish, with the squid crystallin, or with any crystallins from the frog or human lens. Thus, despite the structural similarities between the cubomedusan, squid and vertebrate lenses, their crystallins appear very different.
对两种箱形水母——多手多腕螅(Tripedalia cystophora)和袋状灯水母(Carybdea marsupialis)的透明眼晶状体的超微结构和主要可溶性蛋白质进行了研究。每个物种在四个称为平衡囊的感觉结构上有两只复杂的眼睛(一只大的和一只小的)。晶状体由排列紧密且细胞器较少的细胞组成。晶状体位于视网膜旁边,与光感受器之间仅隔着一层无细胞层。SDS-PAGE显示,袋状灯水母的大晶状体只有两条晶状体蛋白多肽带(分子量约为20,000和35,000道尔顿),而多手多腕螅的有三条带(两条分子量接近20,000道尔顿,一条分子量接近35,000道尔顿)。有趣的是,多手多腕螅的小晶状体似乎明显缺乏或没有低分子量蛋白质。通过FPLC分析,晶状体蛋白表现为单体蛋白,并且在蛋白质免疫印迹中与主要鱿鱼晶状体蛋白、鸡δ-晶状体蛋白或小鼠γ-晶状体蛋白的抗血清没有免疫反应。与抗小鼠α-和β-晶状体蛋白血清的反应非常微弱。多手多腕螅的35,000道尔顿晶状体蛋白被纯化并称为J1-晶状体蛋白。它含有相对较高的亮氨酸(13%)和酪氨酸(9%)以及较低的蛋氨酸(2%)。对几条胰蛋白酶肽段进行了测序。发现与α-和β-晶状体蛋白有微弱的序列相似性,这可能解释了与这些脊椎动物晶状体蛋白明显的微弱免疫交叉反应。用J1-晶状体蛋白的合成肽制备的兔多克隆抗血清在免疫印迹中与多手多腕螅和袋状灯水母的J1-晶状体蛋白强烈反应;相比之下,与这些水母的低分子量晶状体蛋白、鱿鱼晶状体蛋白或青蛙或人类晶状体的任何晶状体蛋白均无反应。因此,尽管箱形水母、鱿鱼和脊椎动物的晶状体在结构上有相似之处,但它们的晶状体蛋白却非常不同。